Automated, high-throughput derivation, characterization and differentiation of induced pluripotent stem cells

被引：0

作者：

Paull, Daniel ^{[1
]}

Sevilla, Ana ^{[1
]}

Zhou, Hongyan ^{[1
]}

Hahn, Aana Kim ^{[1
]}

Kim, Hesed ^{[1
]}

Napolitano, Christopher ^{[1
]}

Tsankov, Alexander ^{[2
,3
,4
]}

Shang, Linshan ^{[1
]}

Krumholz, Katie ^{[1
]}

Jagadeesan, Premlatha ^{[1
]}

Woodard, Chris M. ^{[1
]}

Sun, Bruce ^{[1
]}

Vilboux, Thierry ^{[5
,6
]}

Zimmer, Matthew ^{[1
]}

Forero, Eliana ^{[1
]}

Moroziewicz, Dorota N. ^{[1
]}

Martinez, Hector ^{[1
]}

Malicdan, May Christine V. ^{[5
]}

Weiss, Keren A. ^{[1
]}

Vensand, Lauren B. ^{[1
]}

Dusenberry, Carmen R. ^{[1
]}

Polus, Hannah ^{[1
]}

Sy, Karla Therese L. ^{[1
]}

Kahler, David J. ^{[1
]}

Gahl, William A. ^{[5
,7
,8
]}

Solomon, Susan L. ^{[1
]}

Chang, Stephen ^{[1
]}

Meissner, Alexander ^{[2
,3
,4
]}

Eggan, Kevin ^{[2
,3
,4
,9
]}

Noggle, Scott A. ^{[1
]}

机构：

[1] New York Stem Cell Fdn Res Inst, New York, NY 10023 USA

[2] Broad Inst, Cambridge, MA USA

[3] Harvard Univ, Harvard Stem Cell Inst, Cambridge, MA 02138 USA

[4] Harvard Univ, Dept Stem Cell & Regenerat Biol, Cambridge, MA 02138 USA

[5] NHGRI, Sect Human Biochem Genet, Med Genet Branch, NIH, Bethesda, MD 20892 USA

[6] Inova Hlth Syst, Inova Translat Med Inst, Div Med Genom, Falls Church, VA USA

[7] NIH, NIH Undiagnosed Dis Program, Common Fund, Off Director, Bethesda, MD 20892 USA

[8] NHGRI, NIH, Bethesda, MD 20892 USA

[9] Howard Hughes Med Inst, Cambridge, MA USA

来源：

NATURE METHODS | 2015年 / 12卷 / 09期

基金：

美国国家卫生研究院;

关键词：

EFFICIENT GENERATION; IPSC; MAINTENANCE; INDUCTION; EXPANSION; PLATFORM; EROSION; CULTURE;

D O I：

10.1038/NMETH.3507

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Induced pluripotent stem cells (iPSCSCs) are an essential tool for modeling how causal genetic variants impact cellular function in disease, as well as an emerging source of tissue for regenerative medicine. The preparation of somatic cells, their reprogramming and the subsequent verification of iPSCSC pluripotency are laborious, manual processes limiting the scale and reproducibility of this technology. Here we describe a modular, robotic platform for iPSCSC reprogramming enabling automated, high-throughput conversion of skin biopsies into iPSCSCs and differentiated cells with minimal manual intervention. We demonstrate that automated reprogramming and the pooled selection of polyclonal pluripotent cells results in high-quality, stable iPSCSCs. These lines display less line-to-line variation than either manually produced lines or lines produced through automation followed by single-colony subcloning. The robotic platform we describe will enable the application of iPSCSCs to population-scale biomedical problems including the study of complex genetic diseases and the development of personalized medicines.

引用

页码：885 / +

页数：13

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